This is a protocol preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Method Article
Per-Olof Berggren
Berggren&Caicedo Groups (University of Miami)
Corresponding Author
Alejandro Caicedo
Berggren&Caicedo Groups (University of Miami)
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
An important aspect of studying pancreatic islet physiology is to be able to measure hormone release from islet cells in response to specific stimuli. We demonstrate here step-by-step how to quantitatively measure hormone release from viable isolated pancreatic islets using the in vitro perifusion assay. Islet perifusion has two great advantages over other existing assays: (1) It allows pharmacological manipulation to dissect out signaling mechanisms underlying release of different hormones in the pancreatic islets and, (2) it allows performing simultaneous experiments on multiple islet preparations resulting in high throughput readouts. Other practical applications of islet perifusion include assessment of islet function and viability for experimental purposes and/or clinical application in human islet transplantation. The experimental conditions during islet perifusion are adjusted to simulate the islet “physiological” environment within the pancreas. Therefore, the overall time required to execute the described protocol will be more than 1 h to establish such conditions but should not exceed 2 h to avoid compromising islet health.
Keywords
Perifusion, perifused islet, autocrine, paracrine, signaling, pancreas, islets of Langerhans, acetylcholine, glutamate, GABA
Figure 1
Figure 2
No conflicts disclosed
Loading...
Version History
CURRENT STATUS: Posted
Version 1
Posted 10 Oct, 2011
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Associated Publications
Alpha cells secrete acetylcholine as a non-neuronal paracrine signal priming beta cell function in humans
by Rayner Rodriguez-Diaz, Robin Dando, M Caroline Jacques-Silva, +7
Nature Medicine (08 September, 2011)
ATP-gated P2X3 receptors constitute a positive autocrine signal for insulin release in the human pancreatic cell
by M. C. Jacques-Silva, M. Correa-Medina, O. Cabrera, +11
Proceedings Of The National Academy Of Sciences (06 October, 2011)
Glutamate Is a Positive Autocrine Signal for Glucagon Release
by Over Cabrera, M. Caroline Jacques-Silva, Stephan Speier, +11
Cell Metabolism (06 October, 2011)
Method Article
Per-Olof Berggren
Berggren&Caicedo Groups (University of Miami)
Corresponding Author
Alejandro Caicedo
Berggren&Caicedo Groups (University of Miami)
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 10 Oct, 2011
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
An important aspect of studying pancreatic islet physiology is to be able to measure hormone release from islet cells in response to specific stimuli. We demonstrate here step-by-step how to quantitatively measure hormone release from viable isolated pancreatic islets using the in vitro perifusion assay. Islet perifusion has two great advantages over other existing assays: (1) It allows pharmacological manipulation to dissect out signaling mechanisms underlying release of different hormones in the pancreatic islets and, (2) it allows performing simultaneous experiments on multiple islet preparations resulting in high throughput readouts. Other practical applications of islet perifusion include assessment of islet function and viability for experimental purposes and/or clinical application in human islet transplantation. The experimental conditions during islet perifusion are adjusted to simulate the islet “physiological” environment within the pancreas. Therefore, the overall time required to execute the described protocol will be more than 1 h to establish such conditions but should not exceed 2 h to avoid compromising islet health.
Figure 1
Figure 2
Loading...
Associated Publications
Alpha cells secrete acetylcholine as a non-neuronal paracrine signal priming beta cell function in humans
by Rayner Rodriguez-Diaz, Robin Dando, M Caroline Jacques-Silva, +7
Nature Medicine (08 September, 2011)
ATP-gated P2X3 receptors constitute a positive autocrine signal for insulin release in the human pancreatic cell
by M. C. Jacques-Silva, M. Correa-Medina, O. Cabrera, +11
Proceedings Of The National Academy Of Sciences (06 October, 2011)
Glutamate Is a Positive Autocrine Signal for Glucagon Release
by Over Cabrera, M. Caroline Jacques-Silva, Stephan Speier, +11
Cell Metabolism (06 October, 2011)